Technique Transition

Recombinase polymerase amplification (RPA) is a wellestablished,
versatile isothermal alternative to traditional polymerase chain
reactions (PCR). The development of RPA began with the principles behind
the recombination systems of Escherichia coli and T4-like
bacteriophages in the early 2000s, and the technique is now frequently
used for testing outside the laboratory environment, where the use of
complex and bulky instrumentation – such as thermal cyclers – is
impractical. This flexible technology has recently been turned on its
head with the launch of liquid kits that can be applied across a much
broader range of applications, while still taking advantage of the speed
and sensitivity of the original RPA methodology. In contrast to the
lyophilised formats, that are ideal for low throughput point-of-care
testing, liquid is a good choice for higher throughput lab-based
applications where using very small reaction volumes is more
costeffective. Olaf Piepenburg talks about the progression of RPA and
discusses how the development of liquid technology is opening up new
application opportunities.

The rise of molecular techniques – such as NGS – has caused a
genomics explosion across a wide range of applications, from clinical
and veterinary diagnostics to food and environmental testing. The
mechanisms of these molecular technologies, combined with the demand for
ever-greater sensitivity, mean that relatively large amounts of genetic
material are required to generate high-quality data, making nucleic
acid amplification an essential part of many laboratory workflows. PCR
is considered the gold standard for DNA/RNA amplification and, as a
result, PCR and reverse transcription polymerase chain reaction (RT-PCR)
are now virtually ubiquitous techniques across the life sciences
sector. However, is PCR really the best solution in every situation?

A major advantage of PCR is its simplicity, requiring just a single
enzyme – two for RT-PCR – plus the application-specific primers.
However, this biochemical simplicity comes at the cost of greater
physical complexity. PCR relies on repeatedly heating and cooling
samples to melt the DNA into single strands that can be recognised and
bound by the polymerase enzymes and primers. This means that PCR has
relatively high associated equipment costs and is difficult to perform
outside of the laboratory environment. The need for rapid and efficient
heating and cooling also makes PCR techniques difficult to scale –
larger volume reactions require a significant amount of energy to
rapidly heat and cool, and very low volume reactions require complex
microfluidic systems to avoid evaporation. Finally, the need to heat the
sample to in excess of 90°C to separate the DNA strands will,
obviously, have a significant detrimental impact on any other enzymes or
proteins present in the sample, making PCR incompatible with many other
enzymatic processes. This prevents other enzyme-driven reactions being
run in parallel to the PCR process, significantly limiting the design of
PCR-based assays and workflows.

Dr Olaf Piepenburg is Chief Scientific Officer at TwistDx (now part of Abbott). He was awarded his PhD for work on transcriptional regulation during the embryogenesis of Drosophila at the Max Planck Institute for Biophysical Chemistry in Goettingen, Germany, in 2000. He was a postdoctoral researcher at the Wellcome Institute for Cancer Research and Developmental Biology, Cambridge, UK, before joining TwistDx in 2003, where he co-developed the company’s core RPA technology.

Mauritius Island – An Emerging Centre for R&D in Biotechnology and the Life Sciences

CIDP (Centre International de Développement Pharmaceutique)

Mauritius, the tropical island situated in the Indian Ocean and known
worldwide for its beautiful beaches, is also internationally recognised
for its rule of law, and political and social stability. Over the past
few years, the economy has been successfully transitioned from a
monocrop to a diversified innovation-driven and knowledge-based economy,
resting on agribusiness, export-oriented manufacturing, tourism,
financial services, property development and real estate, ICT-BPO, the
seafood industry, a free port, logistics and a nascent ocean economy.
Emerging sectors such as healthcare and life sciences are presenting
some niche areas for the taking, and the enabling environment is being
put in place to make it happen - especially in the light of sustained
growth within pharmaceutical, medical device, and clinical research.
Important
international players are already in operation locally as the country
has established the appropriate legal and regulatory frameworks based on
international norms, for the development of a strong biomedical
research sector.More info >>